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Fig. 2. Recognition by Rump correlates with nos RNA localization signal
function. (A) UV-crosslinking of MBP-Rump to radiolabeled
nos +1 and +2' element RNA probes. (B) UV-crosslinking
of MBP-Rump to radiolabeled +2' element RNA in the absence (0) or
presence of 200-, 400- and 600-fold molar excess of unlabeled +2' or +1
element RNA. (C) Above is a diagram of the +2' element with the
two CGUU motifs (A and D) indicated. The lightly shaded region is highly
conserved between D. melanogaster and D. virilis
(Bergsten et al., 2001). The
portions of the +2' element retained in the deletion mutants
(
1-5) are indicated by solid bars. C-to-G point mutations in the CGUU
motifs were introduced individually (A or D) or in combination (AD) into the
full-length +2' element. Beneath is shown UV-crosslinking of MBP-Rump to
the indicated radiolabeled RNA probes. (D) Diagram of the
nos-tub:nos+2 transgene, which contains only the nos +1 and
+2' localization elements. (E,F) In situ hybridization to
nos in preblastoderm nosBN embryos carrying (E)
the nos-tub:nos+2 and (F) the nos-tub:nos+2(D)G:C
transgenes. Because nosBN embryos lack endogenous
nos mRNA (Wang et al.,
1994), only the transgenic mRNA is detected.